Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Activation of OGG1 Inhibits Paraptosis in Lens Epithelial Cells of Early Age-Related Cortical Cataract

doi: 10.1167/iovs.66.1.29

Figure Lengend Snippet: Activation of OGG1 inhibits ROS-induced LEC paraptosis. ( A ) Representative immunofluorescence images of indicated LECs against 8-oxoG. ( B ) Quantification and statistical analysis of 8-oxoG fluorescence intensity. ( C , D ) The Western blot images and statistical analysis display the relative protein expression levels of OGG1, normalized to β-actin, after treatment with 50 µM H 2 O 2 for 24 hours. ( E ) Representative confocal images of OGG1-mCherry LECs following treatment at the indicated time points after laser micro-irradiation sites pointed by white arrows . ( F ) Connecting lines of mean values representing OGG1-mCherry recruitment kinetics. ( G , H ) The Western blot images and statistical analysis display the relative protein expression levels of ALIX, GRP78, p-ERK, and p-P38, normalized to β-actin. ( I ) ER and mitochondria were labeled with ER-Tracker green and Mito-Tracker red, respectively, after LECs were treated as indicated. Red arrowheads indicate the dilated ER. ( K ) Quantification and statistical analysis of intracellular vesicles. ( J , L ) Representative images of ROS generation and statistical results of indicated LECs treated with 50 µM H 2 O 2 and 10 µM TH10785. Data are displayed as mean ± SEM from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: TH10785 (MCE), an OGG1 activator, was used at 10 µM for 8 hours.

Techniques: Activation Assay, Immunofluorescence, Fluorescence, Western Blot, Expressing, Irradiation, Labeling

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Activation of OGG1 Inhibits Paraptosis in Lens Epithelial Cells of Early Age-Related Cortical Cataract

doi: 10.1167/iovs.66.1.29

Figure Lengend Snippet: Effects of TH10785 on ROS-induced paraptosis and vacuoles in rat lens. Rat lenses were subjected to control, H 2 O 2 (50 µM), and H 2 O 2 (50 µM) with TH10785 (10 µM) medium for 5 days. ( A , B ) Representative images of cultured rat lenses and the percentage of lens opacity in the control, H 2 O 2 , and H 2 O 2 + TH10785 groups. ( C , D ) TEM showed that TH10785 treatment alleviated the dilated ER in LECs under H 2 O 2 stress. ( E , F ) H&E staining shows that TH10785 treatment reduced the area of vacuoles in the superficial cortex of rat lenses. ( G , H ) Reverse transcription–quantitative real-time PCR was performed to detect the mRNA levels of paraptosis-related genes ( GRP78 and ALIX ) in the LECs. Data are displayed as mean ± SEM from three independent experiments. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: TH10785 (MCE), an OGG1 activator, was used at 10 µM for 8 hours.

Techniques: Control, Cell Culture, Staining, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Activation of OGG1 Inhibits Paraptosis in Lens Epithelial Cells of Early Age-Related Cortical Cataract

doi: 10.1167/iovs.66.1.29

Figure Lengend Snippet: Schematic illustrating the mechanism of ROS-induced paraptosis in LECs of early ARCC patients. This comprehensive mechanism reveals that the elevated ROS orchestrates a cascade of cellular events, including ER stress induction, IGFIR activation, and subsequent mitogen-activated protein kinase pathway signaling. These molecular events result in ER dilation-formed intracellular vesicles and, ultimately, paraptosis in LECs. Notably, TH10785-mediated enhancement of OGG1 activity effectively suppresses ROS-induced paraptosis and vacuolar degeneration in the superficial lens cortex. AC, anterior chamber; C, cornea; CB, ciliary body; I, iris; N, nucleus; PC, posterior chamber; V, vacuoles.

Article Snippet: TH10785 (MCE), an OGG1 activator, was used at 10 µM for 8 hours.

Techniques: Activation Assay, Activity Assay